Low dosed 15-deoxyspergualin preparations

ABSTRACT

15-deoxyspergualin or a pharmaceutically compatible salt thereof is suitable for the preparation of an immunosuppressive, anti-inflammatory or anti-tumoral pharmaceutical, which is effective in a low dosage of 20 to 600 μg/kg body weight when it is applied to the mucous membrane, to which end it is preferably prepared as a dosing aerosol.

This application is a 371 of PCT/EP98/03927 filed Jun. 26, 1998.

DESCRIPTION

The present invention concerns the use of 15-deoxyspergualin for theproduction of an immunosuppressive, anti-inflammatory and anti-tumoralpharmaceutical for administration to the mucous membrane in a dosage inthe range from 20 to 600 μm/kg body weight per day, especially fortreating autoimmune illnesses, inflammatory skin disorders, cancerdiseases and for the prevention of transplant rejection. The activesubstance (±) 15-deoxyspergualin with the formula

    H.sub.2 NC(═NH)NH(CH.sub.2).sub.8 CONHCH(OH)CONH(CH.sub.2).sub.4 NH(CH.sub.2).sub.3 NH.sub.2 (3HCl)

is a synthetic derivative of the anti-tumoral antibiotic spergualin.

The compound can be prepared either as a racemic mixture (cf, forexample, U.S. Pat. No. 4,518,532) or else in optically pure form (cf.,for example, EP-A-0 094 632).

Although numerous publications describe the immunosuppressive propertiesof this compound and its potential suitability for treating autoimmuneillnesses, such as multiple sclerosis, lupus erythematosus and forpreventing transplant rejection, its clinical benefit is very limited onaccount of the very high doses required and the associated side effects.

The main side effect observed in long-term therapy was a temporary dropin leucocyte count with all the consequences of a strongimmunosuppression (cf. Amemiya, H,. et al.: Japanese [Journal] ofTransplantation, 26, 615). The high dosages were regarded as necessarybecause of these demonstrable immunosuppressive effects in the therapyof acute and chronic rejection as well as in the treatment of autoimmuneillnesses (cf. Amemiya., et al.: Transplantation Proceedings XXIII, (1),1087-1089, February 1991).

Conventionally, 15-deoxyspergualin is administered intravenously in adosage of approx. 4 to 6 μg/kg of body weight to obtain animmunosuppressive effect. The effect is characterized by a non-specificgeneral suppression of the immune system.

For treatment of autoimmune illnesses and cancer diseases and for theprevention of transplant rejection, at present no satisfactory long-termtreatments with low side effects are available. The object of theinvention is therefore to make available a preparation for the treatmentof the aforementioned illnesses, which has low side effects, is easy toadminister, does not develop significant side effects even duringlong-term use and is therapeutically effective. Furthermore, thepreparation should be easy to administer even by the patient himself.

Surprisingly, it has now been found that extremely small dosages (20 to600 micrograms/kg body mass) are sufficient to obtain the desiredimmunosuppressive pharmacological effects if the active substance isapplied by way of the respiratory mucous membranes. The risk ofundesired effects is considerably reduced, The ordeal of daily injectionis eliminated. However, the low-dosage therapy presupposes that precisedosage systems are used in order to ensure the necessary constancy ofdosage of highly effective immunosuppressants. In the dosage accordingto the invention, the active ingredient DSG can be administered by meansof the preferred dosing aerosol precisely in the microgram range to thenasal mucous membrane, the respiratory tracts or other mucous membranes,the individual dosage being determined by the amount of active substanceper stroke (200 to 800 microgram per stroke). Because of the low and, atthe same time, precise dosage in the microgram range, it is possible forthe first time to realize in practice the pharmacological effects thatcan be obtained in this dosage. In the process, both local and systemiceffects are produced, since a sufficient amount of the active substanceis resorbed and passes into the bloodstream. For long-term therapy,transdermal systems are also suitable in the low-dosage range.

The object of the invention is the use of 15-deoxyspergualin or apharmaceutically compatible salt thereof as active substance, ifappropriate with the conventional auxiliary and carrier substancesand/or other active substances for the preparation of animmunosuppressive anti-inflammatory or anti-tumoral pharmaceutical inthe form of a preparation intended for application to the mucousmembrane in a dosage of 20 to 600 μg/kg body weight per day, preferablyin the form of a dosing aerosol.

Since the immunosuppressive anti-inflammatory and anti-tumoral effect atthis low dosage is very selective, only slight, if any, side effects,such as, for example, dysaesthesias, are to be expected, even inlong-term therapy.

As a result, autoimmune illnesses, certain cancer diseases andtransplant rejection reactions can be treated with low side effects.According to the invention, autoimmune illnesses of any origin and anyorgan can be treated, for example myasthenia gravis, myositis, multiplesclerosis, Hashimoto's thyroiditis, athropic gastritis, colitisulcerosa, nephrotoxic nephritis, lupus illnesses, arthritis,scleroderma, granulomatosis, autoimmune hepatitis, endo- andmyocarditis, autoimmune cytopenia, psoriasis, neurodermatitis,dermatitis herpetiformis, eczema, morbus basedow, pancreatitis, uveitis,iritis, glomerulonephritis, rheumatoid arthritis, Alzheimer's disease,AIDS and the like. The treatment of multiple sclerosis, lupuserythmatosus, virus-induced autoimmune diseases, AIDS,glomerulonephritis, psoriasis and neurodermatitis is preferred. Thecancer diseases that can be treated include leukaemia and solid tumours,for example, of the lung, breast, sarcoma, ovaries, lymphoma. Transplantrejection reactions may occur in the transplantation of any human organor tissue. The fate of the transplant depends essentially on the natureand extent of the immune reaction in the recipient. As a result of theimmune suppression caused by the pharmaceutical obtained in accordancewith the invention, the survival time of the transplant is considerablyimproved.

Hitherto, the active substance deoxyspergualin could only beadministered intravenously or intramuscularly on account of the highdoses required. The invention, by contrast, allows the active substanceto be administered effectively in the low dosage required in accordancewith the invention by mucous-membrane application.

The essential dosage within the scope of the invention is in the rangefrom 20 to 600 μg/kg body weight per day. It is preferably reliablymaintained therein that the pharmaceutical in accordance with theinvention is packaged and used in a dosing aerosol applicator, which,with each actuation, administers a specified amount of active substance.If, for example, the dosing aerosol device is arranged such that, witheach stroke, aerosol with a content of 500 μg of active substance issprayed out, then, for a patient of 60 kg body weight and a dosage of100 μg/kg body weight per day; this results in 6 pump actions to eachside of the nose, corresponding to 2 pump actions per side of the nose3× per day. The daily dose is suitably administered distributed betweenone to three individual doses.

The dosage to be administered in a particular case will be decided bythe doctor, taking into account the severity of the condition to betreated, the patient's age, sex and weight, and the duration of thetreatment.

Although the pharmaceutical to be produced according to the invention ispreferably formulated as an aerosol, it can also, for example, be in theform of an ointment, a cream, drops, or a lotion, which is formulatedfor application to the mucous membrane, preferably to the nasal mucousmembrane.

Administration to the nasal mucous membrane or respiratory mucousmembrane, for example, in the form of a nasal spray, aerosol, nasalointment or nasal drops is preferred, since the ordeal for the patientis thereby considerably reduced and the patient's compliance can thusalso be improved. As a result of the low dosage of the active substance,such nasal administration is possible for the first time, since, byvirtue of the low concentrations, no irritation of the mucous membraneoccurs. There is also no effect on the ciliar function. Also appropriateis buccal administration, in which the active substance can be resorbedvia the oral mucous membrane. Dosed administration of the pharmaceuticalin accordance with the invention is also possible via the vaginal mucousmembrane and the intestinal mucous membrane, in which case theformulation then assumes the conventional preparation form for thisapplication, for example as a suppository.

The active substance deoxyspergualin can be packaged for use inaccordance with the invention in a manner known per se and processed fora galenic form of application. To this end auxiliary materials andcarriers conventional in the field of pharmaceutical formulations forthe particular form of administration are used.

The active substance can either be used as a free base or as apharmaceutically compatible acid addition salt together withconventional carriers and other conventional pharmaceutical additivesare used. Examples of pharmaceutically compatible acid addition saltswith organic and inorganic acids include, for example, chlorides,hydroxides, lactates, malates, fumarates, acetates, carbonates,citrates, tartrates, glycolates, etc.

15-Deoxyspergualin can either be used as a single active substance orelse together with other active substances as well as vitamins, mineralsand known pharmaceutical additives. Examples of suitable further activesubstances are those with a pharmacological effect, in so far as theyare compatible with 15-deoxyspergualin, for example corticosteroids suchas cortisone, camphor, echinacea, euphorbium etc. or active substanceswith a complementary accompanying effect, such as fumaric acid, activesubstances promoting the absorption and penetration of15-deoxyspergualin, such as 2-propanol or propylene glycol, activesubstances with a soothing effect, such as camomile oil, Azulene,panthenol, retinol palmitate and glycerine.

For the production of a preparation to be administered preferably to therespiratory or nasal mucous membrane, all auxiliary and carriersubstances conventionally used in the field of mucous membrane agentscan be used. The preparation, except for the preferred embodiment asdosing aerosol, can also take any other form appropriate for theprecisely dosable administration via the mucous membrane, and may exist,for example, in the form of a nasal spray, nasal drops, nasal ointment,nasal suspension, nasal gel or the like. To this end base materialsadapted to the form of administration may be used, such as medium-chaintriglycerides, peanut oil, Carbomer, Makrogol, wool wax, alcohol salbenebase, water for injection, paraffin liquidum and excipients such aspreservatives, for example benzalkonium chloride, thiomersal,butylhydroxytoluene, PHB ester, isotonic agents, such as sodiumdihydrogenphosphate or sodium chloride, and flavouring and aromatizingsubstances, such as fennel oil, eucalyptus oil, peppermint oil, dwarfpine-needle oil, cedar-leaf oil, and the like can be used. The nasalspray may take the form of a pump spray or be embodied in a pressurevessel with a pharmacologically acceptable propellant gas. Particularlypreferred is the packaging in a dosing aerosol container.

The combination of deoxyspergualin with magnesium is particularlypreferred. It was found that the compatibility of this combination isparticularly high. Magnesium reliably prevents the (rarely) occurringsubjective complaints (for example, dysaesthesias) during treatment with15-deoxyspergualin.

The invention is described in greater detail below with reference to theexamples.

EXAMPLES Example 1

Nasal Spray

By mixing 2.5 mg and 5 mg respectively of pulverized 15-deoxyspergualinwith 100 g of a carrier substance, e.g. glucose in purified water, anasal spray is prepared in a manner known per se. Application is asdirected by the doctor.

Example 2

Nasal Drops

By mixing 2.5 mg, 5 mg and 10 mg respectively of powdered15-deoxyspergualin with 100 ml of a liquid carrier medium, e.g. glycol,and conventional auxiliaries, nasal drops are prepared in a manner knownper se. Dosage is as directed by the doctor.

Example 3

Ointment

10 mg of 15-deoxyspergualin and 50 g of Eucerin are processed withconventional auxiliaries and additives and processed to prepare anointment. The ointment is applied to the mucous membrane several timesper day.

Example 4

Ointment

10 mg of 15-deoxyspergualin are processed with 50 g of purifiedVaseline, ethereal oils and Azulene in a manner known per se. Theointment is applied to the mucous membrane several times per day.

Example 5

Ointment

10 mg of 15-deoxyspergualin is processed with 50 g of Eucerin and 1.0 gof fumaric acid, if appropriate with the addition of fragrances andconservation agents, to prepare an ointment. The ointment is applied tothe mucous membrane several times per day.

Example 6

Nasal Oil

250 mg of deoxyspergualin are processed with paraffinum liquidum,benzalkonium chloride and sodium citrate in a manner known per se toprepare 10 ml of nasal oil. The nasal oil is introduced into the noseaccording to the dosage directions of the doctor.

Example 7

Nasal Drops

20 mg of deoxyspergualin are processed with 2 mmol of magnesium chlorideand 100 ml of purified water, if appropriate with the addition ofcamomile flower fluid extract, in a manner known per se. Dosage iscarried out as directed by the doctor.

Example 8

Dosing Aerosol

50 mg of deoxyspergualin are processed with ethyl acetate, sorbitoltrioleate, dichlorodifluoromethane and trichlorofluoromethane to form adosing aerosol comprising 200 individual doses. Dosage is carried out asdirected by the doctor.

Example 9

Treatment of Multiple Sclerosis Relapsing Form (EDSS<5).

Patient 1: female, 31 years, 60 kg body weight, MS diagnosis 1989 byclinic, magnetic resonance imaging and evidence of oligoclonal bandingin cerebrospinal fluid, so far approximately 2 relapses per year, slowlyprogressive incapacity, presentation August 1994, walking range approx.100 m with stick, slight paraspastis, dynaesthesias, pensioned 12 monthsago.

Therapy was carried out with 150 μg/kg body weight of 15-deoxyspergualinas aerosol, applied to the nasal mucous membrane over 14 days. Onfurther presentation after 6 weeks, the walking range was now approx.half an hour. There were no more dysaesthesias and paraspastis. The gaitwas normal. Further therapy with 15-deoxyspergualin over 14 days led toa normal condition which has remained unchanged since. Since June 1997working again, no new relapses. Neither subjective nor objective sideeffects of the therapy were observed.

Patient 2: male, 25 years, 81 kg body weight, MS diagnosis 1992 byclinic, magnetic resonance imaging and evidence of oligoclonal bandingin cerebrospinal fluid, so far approximately 4 relapses per year, slowlyprogressive incapacity, presentation July 1995, walking range approx. 50m, ataxis, ataxic-spastic gait, intense tiredness.

A nasal-aerosol therapy with 150 μg/kg body weight of 15-deoxyspergualinover 14 days produced a significant improvement at the next presentationafter 7 weeks. The tiredness had disappeared, the walking range hadincreased to about 5 km and the gait had normalized. A further 14-daytherapy with 150 μg/kg body weight per day 15-deoxyspergualin produced afurther improvement. No further relapses occurred. Overall, normalmobility is now present. Neither subjective nor objective side effectsof the therapy were observed.

Example 10

Treatment of Multiple Sclerosis, Primary Chronic Progressive Form

Patient 1: male, 29 years, 65 kg, MS diagnosis in 1991 by clinic,magnetic resonance imaging and oligoclonal banding in cerebrospinalfluid, presentation March 1996, walking range approx. 100 m,spastic--ataxic gait, disturbed fine motor skill, intense tiredness,incapable of working (student).

A 16-day aerosol therapy with 100 μg/kg body weight of15-deoxyspergualin per day brought about a significant improvement. Thetiredness had disappeared, the walking range had increased to about 500m, the fine motor skill was normal again and the gait had visiblyimproved. The patient felt able to meet his demands again. Furthertherapy with 15-deoxyspergualin over 16 days brought about a furtherstabilization of the general condition, and the patient felt furthercapable of meeting his demands. Neither subjective nor objective sideeffects of the therapy were observed.

Patient 2: Female, 41 years, 57 kg, MS diagnosis in 1987 by clinic,magnetic resonance imaging and oligoclonal banding in cerebrospinalfluid, presentation May 1996, chronic tiredness, walking range 150 m,spastic-ataxic gait, significantly reduced strength in hands, slightataxia of hands.

After therapy with 100 μg/kg body weight of 15-deoxyspergualin for 18days by infusion, at the next presentation after 6 weeks, the walkingrange had increased to approx. 200 m, the gait was more regular, thestrength of the hands was increased, the fine motor skill was better.Further therapy with 7.5 mg of 15-deoxyspergualin over 18 days as nasalspray stabilized the condition further, so that the patient felt capableof doing her housework again.

Example 11

Treatment of Lupus Erythematosus

a. Lupus Erythematosus (LE), Cutaneous Forn

Patient: male, 12 years, 40 kg, diagnosis in 1995 by skin biopsy andclinic, presentation in July 1996, numerous skin spots and lesions onthe entire body, previous therapy 40 mg Decortin/day, chronic tiredness.

Therapy with 50 μg/kg 15-deoxyspergualin over 20 days by aerosol broughta clear improvement at the further presentation 6 weeks later. The skinlesions and tiredness had disappeared and Decortin could be discontinued. A further 20 days' therapy with 2 mg 15-deoxyspergualin per day bynasal spray brought about a stabilization of the condition. Side effectsof the therapy were not observed.

b. Lupus Erythematosus (LE), Systemic LE (SLE)

Patient: male, 16 years, 42 kg, diagnosis in 1989 by clinic andlaboratory tests, presentation in July 1994, nephrotic syndrome withhigh protein loss (7.5 g/l) from the kidney, butterfly erythem in face,chronic tiredness, school attendance not possible for two years.

A therapy with 250 μg/kg 15-deoxyspergualin per day for 14 days byaerosol brought about a clear improvement in the condition of theillness at the next presentation 5 weeks later. The patient was able toattend school again, the tiredness had disappeared, the proteinexcretion through the kidney (2.9 g/l) was significantly reduced, thebutterfly erythem had faded. A further therapy with 0.25 mg/kg bodyweight of 15-deoxyspergualin over 16 days by infusion brought about afurther improvement. There was no more protein excretion through thekidney, the patient's condition was stable, regular school attendancewas possible.

Example 12

Treatment of Glomerulonephritis

Patient: male, 29 years, 74 kg, diagnosis in 1995 by kidney biopsis,chronic tiredness, presentation November 1995, tiredness, elevatedcreatinine (1.9 mg), liquid retention.

Therapy with 150 μg/kg body weight of 15-deoxyspergualin over 18 days byaerosol brought about a clearly improved condition of the illness at afurther presentation after 6 weeks. The creatinine content in the serumwas reduced (1.3 mg). A further therapy with 150 μg/kg body weight of15-deoxyspergualin over 18 days by infusion brought about a furthernormalization of the creatinine level (0.7 mg). The kidney biopsybrought about a clear reduction in the symptoms of glomerulonephritis.Neither subjective nor objective side effects of the therapy wereobserved.

Example 13

Treatment of Rheumatoid Arthritis (ra)

Patient: male, 36 years, 79 kg, diagnosis in 1987 by clinic and positiverheumafactor, presentation in June 1996, chronic tiredness, painfullimited movement in the finger, knee and shoulder joints.

A therapy with 200 μg/kg body weight of 15-deoxyspergualin over 18 daysby aerosol brought about a dear improvement in the condition of theillness at a further presentation after 6 weeks. The tiredness haddisappeared. The movement in the joints had improved, as had the pains.A further therapy with 7.5 mg 15-deoxyspergualin per day by a nasalspray over 20 days brought about a further stabilization of thecondition of the illness. No further relapses in the illness occurred,the rheumafactor was negative. Neither subjective nor objective sideeffects of the therapy were observed.

Example 14

Treatment of an HIV Infection

Patient: male, 29 years, 72 kg, with confirmed HIV infection anddepressed suppressor cell count (<750) no symptoms of AIDS, presentationin April 1995, chronic tiredness.

A therapy with 7.5 mg 15-deoxyspergualin per day, administered via anasal spray, for five times 18 days improved the tiredness and led to anincrease in CD8 cells to a count of over 900. Neither subjective norobjective side effects were observed.

Example 15

Treatment of Neurodermatitis

Patient: male, 10 years, 22 kg with neurodermatitis, diagnosis 1990,presentation August 1995, painful itching patches on skin on the trunkand extremities, improvement with cortisone ointment.

Therapy with grease-free 15-DSG emulsion (10 mg/10 g emulsion) appliedto the nasal mucous membrane over 20 days. After two repetitions atintervals of 6 weeks, healing of the skin patches without complications,no occurrence of new itching patches. Neither subjective nor objectiveside effects were observed.

Example 16

Treatment of Myasthenia Gravis

Patient: female, 39 years, 62 kg, diagnosis in 1983 by clinic andserology, therapy with 150 μg/kg body weight of 15-deoxyspergualinaerosol for two times 14 days. A lasting neurological improvement withan absence of further relapses occurred. Neither subjective norobjective side effects were observed.

Example 17

Treatment of Leukaemia

Patient: male, 19 years, ALL, diagnosis in 1995 by clinic, diagnosis1995 confirmed by clinic, blood picture, immunohistochemistry and bloodmarrow biopsy, presentation May 1996.

Therapy with 50 μg/kg body weight of 15-deoxyspergualin for 25 days andfurther therapy with 5 mg 15-deoxyspergualin by nasal spray broughtabout a complete remission after 8 weeks until now. Neither subjectivenor objective side effects were observed.

What is claimed is:
 1. Use of 15-deoxyspergualin or a pharmaceuticallycompatible salt thereof as active substance, if appropriate togetherwith conventional auxiliary and carrier substances and/or further activesubstances for the preparation of an immunosuppressive anti-inflammatoryor anti-tumoral pharmaceutical in the form of a preparation intended forapplication to the mucous membrane in a dosage of 20 to 600 μg/kg bodyweight per day.
 2. Use according to claim 1,characterized in thatsaidpharmaceutical is prepared as a dosing aerosol.
 3. Method according toclaim 1,characterized in thatapplication is to the mucous membrane ofthe nose or the respiratory tracts.
 4. Use according to claim 1 for thetreatment of autoimmune illnesses, cancer diseases or transplantrejection reactions.
 5. Use according to claim 4 for the treatment ofmultiple sclerosis, myasthenia gravis, lupus erythematosus,virus-induced autoimmune illnesses, AIDS, glomerulonephritis orpsoriasis, neurodermatitis, eczema.
 6. Use according to claim1characterized in thatthe pharmaceutical additionally contains magnesiumand/or a corticosteroid.
 7. Use according to claim 1 for the preparationof a pharmaceutical as dosing aerosol.
 8. Method for obtainingimmunosuppression by application of a deoxyspergualin or apharmaceutically compatible salt thereof as active substance-containingaerosol in a dosage of 20 to 600 μg/kg body weight per day to the mucousmembrane.
 9. Method according to claim 8,characterized in thatanautoimmune illness, inflammatory illness, cancer disease or transplantrejection reaction is treated.
 10. Method according to claim9,characterized in thatmultiple sclerosis, lupus erythematosus, avirus-induced autoimmune illness, AIDS, glomerulonephritis or psoriasis,neurodermatitis, eczema, myasthenia gravis is treated.
 11. Dosingaerosol device,characterized in thatit contains a pharmaceutical with15-deoxyspergualin or a pharmaceutically compatible salt thereof asactive substance, if appropriate together with conventional auxiliariesand carriers and/or further active substances for the preparation of animmunosuppressive anti-inflammatory or anti-tumoral pharmaceutical whichis formulated for application to the mucous membrane and contains theactive substance in a concentration such that an amount of 20 to 600μg/kg body weight per day is dispensed in the aerosol duringconventional use of device.